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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV140081	2/2	Homo sapiens	NAY	Filtration Microfluidics	Im H	2014	56%
Study summary Full title Label-free detection and molecular profiling of exosomes with a nano-plasmonic sensor All authors Im H, Shao H, Park YI, Peterson VM, Castro CM, Weissleder R, Lee H Journal Nat Biotechnol Abstract Exosomes show potential for cancer diagnostics because they transport molecular contents of the cell (show more...)Exosomes show potential for cancer diagnostics because they transport molecular contents of the cells from which they originate. Detection and molecular profiling of exosomes is technically challenging and often requires extensive sample purification and labeling. Here we describe a label-free, high-throughput approach for quantitative analysis of exosomes. Our nano-plasmonic exosome (nPLEX) assay is based on transmission surface plasmon resonance through periodic nanohole arrays. Each array is functionalized with antibodies to enable profiling of exosome surface proteins and proteins present in exosome lysates. We show that this approach offers improved sensitivity over previous methods, enables portable operation when integrated with miniaturized optics and allows retrieval of exosomes for further study. Using nPLEX to analyze ascites samples from ovarian cancer patients, we find that exosomes derived from ovarian cancer cells can be identified by their expression of CD24 and EpCAM, suggesting the potential of exosomes for diagnostics. (hide) EV-METRIC 56% (90th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin NAY Focus vesicles exosomes Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture Filtration Microfluidics Protein markers EV: Flotilin1/ CD63/ HSP90/ Flotillin2/ HSP70/ CD9 non-EV: Integrin-beta1/ Integrin-alpha5 Proteomics no Show all info Study aim Technical Sample Species Homo sapiens Sample Type Cell culture supernatant EV-harvesting Medium EV Depleted Separation Method Filtration steps 0.22µm or 0.2µm Other Name other separation method Microfluidics Characterization: Protein analysis Western Blot Antibody details provided? No Detected EV-associated proteins CD63/ CD9/ Flotilin1/ HSP90/ HSP70/ Flotillin2 Detected contaminants Integrin-beta1/ Integrin-alpha5 ELISA Antibody details provided? No Detected EV-associated proteins CD63/ CD9/ Flotilin1/ HSP90/ HSP70/ Flotillin2 Detected contaminants Integrin-beta1/ Integrin-alpha5 Characterization: Particle analysis NTA EM EM-type transmission EM/ scanning EM Image type Close-up, Wide-field Report size (nm) Not reported

	EV140081	1/2	Homo sapiens	Ascites	Filtration Microfluidics	Im H	2014	0%
Study summary Full title Label-free detection and molecular profiling of exosomes with a nano-plasmonic sensor All authors Im H, Shao H, Park YI, Peterson VM, Castro CM, Weissleder R, Lee H Journal Nat Biotechnol Abstract Exosomes show potential for cancer diagnostics because they transport molecular contents of the cell (show more...)Exosomes show potential for cancer diagnostics because they transport molecular contents of the cells from which they originate. Detection and molecular profiling of exosomes is technically challenging and often requires extensive sample purification and labeling. Here we describe a label-free, high-throughput approach for quantitative analysis of exosomes. Our nano-plasmonic exosome (nPLEX) assay is based on transmission surface plasmon resonance through periodic nanohole arrays. Each array is functionalized with antibodies to enable profiling of exosome surface proteins and proteins present in exosome lysates. We show that this approach offers improved sensitivity over previous methods, enables portable operation when integrated with miniaturized optics and allows retrieval of exosomes for further study. Using nPLEX to analyze ascites samples from ovarian cancer patients, we find that exosomes derived from ovarian cancer cells can be identified by their expression of CD24 and EpCAM, suggesting the potential of exosomes for diagnostics. (hide) EV-METRIC 0% (median: 13% of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Ascites Sample origin NAY Focus vesicles exosomes Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture Filtration Microfluidics Protein markers EV: non-EV: Proteomics no Show all info Study aim Technical Sample Species Homo sapiens Sample Type Ascites Separation Method Filtration steps 0.22µm or 0.2µm Other Name other separation method Microfluidics Characterization: Particle analysis NTA

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EV-TRACK ID	EV140081
species	Homo sapiens
sample type	Cell culture	Ascites
cell type	NAY	NA
medium	EV Depleted
condition	NAY	NAY
separation protocol	Filtration Microfluidics	Filtration Microfluidics
Exp. nr.	2	1
EV-METRIC %	56	0